RNAseq from P4 murine whole kidneys with Polycystic Kidney Disease

Snap frozen Postnatal day4 murine kidney tissues were crushed using micropestles in RLT Buffer and RNA was extracted from these tissues using QiaShredders followed by Qiagen RNAeasy Mini Kits according to manufacturer’s protocol. RNA concentration and integrity were determined by Aligent bioanalyzer. cDNA library preparation was carried out using NEB Next Ultra Directional RNA Library Prep Kit for Illumina RNA sequencing (E7420S) and sequenced on an Illumina HiSeqX10 machine in 150 bp paired end format. Sequence data was processed with Skewer adaptor trimmer and mapped with HiSat2 to the Mus musculus GRCm38_v90 genome assembly. The resulting Bam files are provided here.Samples were from Control Cre (Hoxb7-cre) mice, Aurka flox/flox; Hoxb7-cre (Aurka Knockout Out -AKO) mice, Inpp5e flox/flox; Hoxb7-cre (Inpp5e Knockout Out -IKO) mice, Pkd1 flox/flox; Hoxb7-cre (Pkd1 Knockout Out -PKO) mice, Inpp5e flox/flox; Aurka flox/flox; Hoxb7-cre (Inpp5e and Aurka double Knockout Out -dKO-I) mice, and Pkd1 flox/flox; Aurka flox/flox; Hoxb7-cre (Pkd1 and Aurka double Knockout Out -dKO-P) mice.Hoxb7-cre mediated deletion of floxed alleles specifically in the kidney collecting ducts.Inpp5e KO results in Polycystic Kidney Disease related to Joubert Syndrome.Pkd1 KO results in Autosomal Dominant Polycystic Kidney disease.Aurka KO had no phenotype and appeared normal.Hoxb7-cre control animals had no phenotype and appeared normal.Both Inpp5e and Aurka, and Pkd1 and Aurka double knockout mice exhibited a rescue in disease.Sample labels abbreviated to groups as bolded above.

本研究采集了产后第4天的鼠肾组织，采用微研杵在RLT缓冲液中将其捣碎，随后利用QiaShredders提取RNA，并按照制造商的方案采用Qiagen RNAeasy Mini试剂盒进行纯化。通过Aligent生物分析仪测定RNA的浓度和完整性。cDNA文库的构建采用NEB Next Ultra定向RNA文库构建试剂盒（E7420S）进行，并在Illumina HiSeqX10测序仪上以150 bp双端测序模式进行测序。序列数据经过Skewer适配器修剪和HiSat2比对至Mus musculus GRCm38_v90基因组组装。所生成的Bam文件已在此提供。样本来自对照组Cre（Hoxb7-cre）小鼠、Aurka flox/flox; Hoxb7-cre（Aurka敲除-KO）小鼠、Inpp5e flox/flox; Hoxb7-cre（Inpp5e敲除-KO）小鼠、Pkd1 flox/flox; Hoxb7-cre（Pkd1敲除-KO）小鼠、Inpp5e flox/flox; Aurka flox/flox; Hoxb7-cre（Inpp5e和Aurka双重敲除-KO-I）小鼠以及Pkd1 flox/flox; Aurka flox/flox; Hoxb7-cre（Pkd1和Aurka双重敲除-KO-P）小鼠。Hoxb7-cre介导的floxed等位基因的特异性删除发生在肾集合管中。Inpp5e敲除导致与Joubert综合征相关的多囊肾病。Pkd1敲除导致常染色体显性多囊肾病。Aurka敲除无表型且外观正常。Hoxb7-cre对照组动物无表型且外观正常。Inpp5e和Aurka以及Pkd1和Aurka双重敲除小鼠显示出疾病缓解。样本标签已缩写为上述加粗的组别。